Abstract 2031: High plex proteomic prognostic marker discovery for patients with pancreatic adenocarcinoma using digital spatial profiling
Authors: Assya Legrini, Holly Leslie, Stephan Dreyer, Joanne Edwards, Andrew Biankin, David Chang, Nigel Jamieson
Journal: Cancer Research (2022)

Abstract Pancreatic Ductal Adenocarcinoma (PDAC) long-term outcomes following surgical resection remain poor, with only 20% of patients surviving 5 years post pancreatectomy. With its immune-privileged nature, starting from the early pre-neoplastic state, it appears to easily evade the antitumor immune response. Although there is rationale for targeting PDAC immune pathways, little benefit has been observed to date. The aim of the study was to interrogate the immune landscape of PDAC utilising the Nanostring GeoMx Digital Spatial Profiler (DSP), enabling high-plex proteomic characterisation whilst maintaining tumor microenvironment (TME) topographical features. We assessed tumor samples from 28 treatment-naive PDAC cases represented in a multiregional FFPE tissue microarray (TMA) for which extensive IHC, molecular, genomic characterisation and clinicopathological follow-up data is available. Multiplex immunofluorescent staining for nuclei, panCK, aSMA and CD3 was used to guide region selection using the GeoMx DSP system in multiple TMA cores. 60 immune markers were quantified simultaneously in different tissue compartments defined by immunofluorescence co-localization including tumor (panCK+), immune stromal (PanCK-) regions. The spatially informed assessment by DSP was validated by both regression and variable prognostication compared with IHC for stromal CD3, CD8 and CD68 in near serial TMA PDAC sections. Unsupervised analysis of proteome data (DSP) in the panCK-negative regions identified an immune poor group associated with shorter OS (13.0 versus 31.1mths, P=0.005). When transcriptomic subtype was considered, the checkpoint inhibitor B7-H3 was significantly upregulated in the squamous subtype tumours versus the classical group (Log2: 1.63, P = 0.001). Patients with high B7-H3 expression, using a median expression cut-off, were associated with shorter OS on multivariate analysis (HR: 4.16, P=0.01) a finding that was validated in an external cohort at the bulk transcriptome level. This pilot scale discovery study shows the potential of the Nanostring DSP technology to identify spatially-informed biomarkers with prognostic relevance in biopsy sized samples from treatment-naive PDAC. A number of relevant candidate immune predictors were identified in a spatial context, which are currently undergoing validation in a larger independent cohort and within the neoadjuvant setting. Future studies will apply this technology to pre and post treatment biopsy samples. Citation Format: Assya Legrini, Holly Leslie, Stephan Dreyer, Joanne Edwards, Andrew Biankin, David Chang, Nigel Jamieson. High plex proteomic prognostic marker discovery for patients with pancreatic adenocarcinoma using digital spatial profiling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2031.